Total protein was extracted from CRC tissues and cultured cells and subjected to immunoblotting, as previously described [20 (link)]. Primary antibodies including rabbit polyclonal anti-claudin-1 (1:1000), anti-claudin-2 (1:500), anti-claudin-3 (1:1000), and anti-claudin-7 (1:1000) were obtained from Abcam (Cambridge, MA, USA). Primary antibody rabbit polyclonal anti-occludin (1:400) was purchased from Invitrogen (Carlsbad, CA, USA). Primary antibodies including rabbit polyclonal anti-E-cadherin (1:1000), anti-c-kit (1:1000), anti-c-Jun (1:1000), anti-p-c-Jun (1:1000), anti-JNK (1:1000), and anti-p-JNK (1:1000) were obtained from Cell Signaling Technology (Beverly, MA, USA). Mouse monoclonal anti-β-actin (1:2000) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The proteins were detected using enhanced chemiluminescence (ECL) (ThermoFisher Scientific, Waltham, MA, USA) and viewed in Fusion FX Vilber Lourmat (Paris, France).
Anti c kit
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-c-Kit is a primary antibody that detects the c-Kit protein, also known as CD117. The c-Kit protein is a receptor tyrosine kinase that plays a crucial role in the regulation of cell growth, survival, and differentiation. This antibody can be used in various research applications, such as western blotting, immunohistochemistry, and flow cytometry, to study the expression and localization of the c-Kit protein in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti stro 1, by R&D Systems (3 mentions)
Anti vimentin, by Abcam (3 mentions)
Anti β actin, by Santa Cruz Biotechnology (2 mentions)
Anti nrf2, by Cell Signaling Technology (2 mentions)
Anti ck14, by Cell Signaling Technology (2 mentions)
Anti 8 ohdg, by Merck Group (2 mentions)
Anti collagen 1 col1, by Abcam (2 mentions)
Anti osteopontin opn, by Abcam (2 mentions)
Anti β actin, by Merck Group (2 mentions)
Anti claudin 1, by Abcam (1 mentions)
Anti e cadherin, by Cell Signaling Technology (1 mentions)
Anti claudin 3, by Abcam (1 mentions)
Anti claudin 7, by Abcam (1 mentions)
Anti p jnk, by Cell Signaling Technology (1 mentions)
Anti p c jun, by Cell Signaling Technology (1 mentions)
Anti occludin, by Thermo Fisher Scientific (1 mentions)
Anti jnk, by Cell Signaling Technology (1 mentions)
Anti claudin 2, by Abcam (1 mentions)
Anti c jun, by Cell Signaling Technology (1 mentions)
Anti p38 mapk, by Abcam (1 mentions)
8 protocols using anti c kit
1
Immunoblotting Analysis of Tight Junction Proteins
2
Protein Expression Analysis of Transmembrane Proteins in Small Intestine
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After feeding the mice GSP (0.5 g/kg) once a day for 5 days, the small intestine samples were collected and prepared by incubating in RIPA buffer. The total protein extracted and an equal amount of protein from the samples were separated using SDS‑PAGE and then transferred to polyvinylidene difluoride membranes. The membranes were probed with the indicated antibodies. Anti‑transmembrane protein 16A (TMEM16A; Abcam, Cambridge, UK), anti‑c‑kit (Cell Signaling Technology, Denver, MA, USA), anti‑transient receptor potential melastatin 7 (TRPM7; Abcam, Cambridge, UK), and anti‑β‑actin (Santa Cruz Biotechnology, Dallas, TX) antibodies were used. All other procedures were conducted as previously described 12 (link).
3
Comprehensive Immunostaining Protocol for Stem Cell Analysis
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Following a 15-min fixation in 4% paraformaldehyde, the samples were permeabilized with 0.1% Triton X-100 for 20 min. Each sample underwent treatment with a blocking agent and subsequent incubation with primary antibodies. The primary antibodies used were as follows: anti-CK14 (1:100; Cell Signaling Technology, Danvers, MA, USA), anti-c-Kit (1:400; Cell Signaling Technology), anti-STRO-1 (1:100; R&D Systems, Minneapolis, MN, USA), anti-Vimentin (1:500; Abcam, Cambridge, UK), anti-8-OHdG (1:100; Sigma), anti-collagen I (COL1; 1:100, Abcam), anti-osteopontin (OPN; 1:100, Abcam), and anti-NRF2 (1:100; Cell Signaling Technology). Following the retrieval of the antibodies, DAPI and the appropriate secondary antibody were incubated. Immunofluorescence images were acquired using a confocal microscope.
4
Comprehensive Antibody Panel for Signaling Pathways
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The following antibodies were used for western blot (WB) and immunofluorescence (IF): anti-α-Tubulin (WB 1:10000, Abcam ref.: ab7291), anti-β-Actin (WB 1:20000, Sigma-Aldrich ref.: A5441), anti-Phospho p38MAPK (T180/Y182) (WB 1:1000, Cell Signaling ref.: #4511), anti-p38MAPK (WB 1:1000, Abcam ref.: ab170099), anti-Phospho VEGFR2 (Y1175) (WB 1:1000, Cell Signaling ref.: #2478), anti-VEGFR2 (WB 1:1000, Cell Signaling ref.: #3770), anti-Phospho c-Kit (Y719) (WB 1:1000, Cell Signaling ref.: #3391), anti-c-Kit (WB 1:1000, Cell Signaling ref.: #3074 and WB 1:1000, Santa Cruz ref.: sc-13508), anti-DUSP2 (WB 1:1000, IF 1:100, Sigma-Aldrich ref.: SAB4300841), PathScan® RTK Signaling Antibody Array Kit (Chemiluminescent Readout, Cell Signaling ref.: #7982).
5
Immunophenotyping of hDPSCs using Confocal Microscopy
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Passage 3 hDPSCs were seeded in 24-well plates for routine culture. When cells reached approximately 80% confluence, the medium was discarded, cells were rinsed and subsequently fixed with 4% of Paraformaldehyde (Solarbio, Zhengzhou, China) at room temperature. The following primary antibodies were added to the cells at the indicated dilution: anti-STRO-1 (1:100; R&D Systems, Minneapolis, MN, USA), anti-Vimentin (1:500; Abcam, Cambridge, UK), anti-Nestin (1:100; Santa Cruz Biotechnology, Houston, TX, USA), anti-c-Kit (1:400; Cell Signaling Technology, Danvers, MA, USA), anti-RUNX2 (1:100; Santa Cruz Biotechnology), and anti-ALP (1:100; Abcam). After a washing step, the cell nuclei were counter-stained with 4′,6-Diamidino-2′-phenylindole (DAPI, Invitrogen, Carlsbad, CA, USA). Cells were analyzed under a confocal microscope (LSM800, Zeiss, Oberkochen, Germany) and the mean fluorescence intensity was determined using the ZEN 2.4 software.
6
FDA-Approved Drug Library Evaluation
7
Comprehensive Immunostaining Protocol for Stem Cell Analysis
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Following a 15-min fixation in 4% paraformaldehyde, the samples were permeabilized with 0.1% Triton X-100 for 20 min. Each sample underwent treatment with a blocking agent and subsequent incubation with primary antibodies. The primary antibodies used were as follows: anti-CK14 (1:100; Cell Signaling Technology, Danvers, MA, USA), anti-c-Kit (1:400; Cell Signaling Technology), anti-STRO-1 (1:100; R&D Systems, Minneapolis, MN, USA), anti-Vimentin (1:500; Abcam, Cambridge, UK), anti-8-OHdG (1:100; Sigma), anti-collagen I (COL1; 1:100, Abcam), anti-osteopontin (OPN; 1:100, Abcam), and anti-NRF2 (1:100; Cell Signaling Technology). Following the retrieval of the antibodies, DAPI and the appropriate secondary antibody were incubated. Immunofluorescence images were acquired using a confocal microscope.
8
Immunofluorescence Staining of Tissue Sections
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After deparaffinization and rehydration, sections were preincubated with 3% bovine serum albumin. These sections were reacted sequentially with the an anti-GDA, anti-PDZK1, or anti-URAT1 antibody and 1:200 Alexa Fluor-labeled goat anti-mouse IgG (488; Molecular Probes, Eugene, OR, USA), with anti-ABCG2 and Alexa Fluor-labeled goat anti-mouse IgG (594; Molecular Probes), or with anti-cKit (Cell signaling technology), anti-uric acid (Santa Cruz Biotechnology) antibody, and Alexa Fluor-labeled goat anti-rabbit IgG (594; Molecular Probes). Nuclei were counterstained with Hoechst 33258 (Sigma-Aldrich). Fluorescence images were evaluated using an image analysis system (Dp Manager 2.1; Olympus Optical Co., Tokyo, Japan) and a Wright Cell Imaging Facility (WCIF) ImageJ software 1.53e (http://www.uhnresearch.ca/facilities/wcif/imagej accessed on 9 August 2021).
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